Primary Cardiomyocyte Hypertrophy Analysis

In our quest to assess cellular hypertrophy, we adopted the RP staining method. In the initial phase, primary cardiomyocytes underwent fixation using a 4% paraformaldehyde solution, after which we permeabilized them with 0.1% Triton. Following this, we proceeded to stain these cells with phalloidin–rhodamine at a dilution ratio of 1:200. In the concluding steps of the procedure, we used 4′,6-diamidino-2-phenylindole (DAPI; Beyotime, Shanghai, China) for counterstaining. We meticulously examined the stained cells and captured their images using a Nikon 80i fluorescence microscope (Nikon, Tokyo, Japan), thus enabling us to closely observe and analyze cellular hypertrophy in these primary cardiomyocytes.